U.S. patent application number 15/257143 was filed with the patent office on 2017-03-09 for vlp stabilized vaccine compositions.
This patent application is currently assigned to Inventprise, LLC. The applicant listed for this patent is Inventprise, LLC. Invention is credited to Subhash V. Kapre.
Application Number | 20170065704 15/257143 |
Document ID | / |
Family ID | 58188631 |
Filed Date | 2017-03-09 |
United States Patent
Application |
20170065704 |
Kind Code |
A1 |
Kapre; Subhash V. |
March 9, 2017 |
VLP Stabilized Vaccine Compositions
Abstract
The invention is directed to compositions and methods for the
stabilization of viral and bacterial vaccines. Vaccines of the
invention are contained in VLPs with stabilizing agents such as,
for example, sugar alcohols (e.g., sorbitol) and degraded gelatins.
Preferably the gelatin has an average molecular weight of 10,000
kilodaltons or less. These vaccines have a substantially improved
thermostability as well as long term stability. The invention is
also directed to the manufacture of a vaccine or the invention and
methods for the administration of a vaccine of the invention to
patients.
Inventors: |
Kapre; Subhash V.; (Redmond,
WA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Inventprise, LLC |
Redmond |
WA |
US |
|
|
Assignee: |
Inventprise, LLC
Redmond
WA
|
Family ID: |
58188631 |
Appl. No.: |
15/257143 |
Filed: |
September 6, 2016 |
Related U.S. Patent Documents
|
|
|
|
|
|
Application
Number |
Filing Date |
Patent Number |
|
|
62214526 |
Sep 4, 2015 |
|
|
|
Current U.S.
Class: |
1/1 |
Current CPC
Class: |
C12N 2710/20034
20130101; C12N 2730/10123 20130101; A61K 2039/5258 20130101; C12N
2730/10142 20130101; A61K 47/42 20130101; A61K 9/0019 20130101;
A61K 47/10 20130101; C12N 7/00 20130101; A61K 9/19 20130101; A61K
39/12 20130101; C07K 14/005 20130101; A61P 31/12 20180101 |
International
Class: |
A61K 39/12 20060101
A61K039/12; C07K 14/005 20060101 C07K014/005; C12N 7/00 20060101
C12N007/00; A61K 9/19 20060101 A61K009/19; A61K 47/42 20060101
A61K047/42; A61K 47/10 20060101 A61K047/10 |
Claims
1. A vaccine comprising VLPs that encapsulate an immunogenic
component and a stabilizing agent.
2. The vaccine of claim 1, wherein the VLPs are derived from the
structural components of a virus.
3. The vaccine of claim 2, wherein the VLPs are derived from
hepatitis virus or human papilloma virus.
4. The vaccine of claim 1, wherein the immunogenic component
comprises an immunogenic portion of a virus, a bacterium, a
parasite or a combination thereof.
5. The vaccine of claim 1, wherein the stabilizing agent comprises
one or more gelatins, one or more amino acids, or one or more sugar
alcohols, or any combination thereof.
6. The vaccine of claim 5, wherein the one or more gelatins are
degraded chemically or mechanically.
7. The vaccine of claim 6, wherein the one or more degraded
gelatins are degraded to an average molecular weight of ten
kilodaltons or less.
8. The vaccine of claim 6, wherein the one or more degraded
gelatins are degraded to an average molecular weight of eight
kilodaltons or less.
9. The vaccine of claim 1, wherein the stabilizing agent comprises
a sugar alcohol.
10. The vaccine of claim 9, wherein the sugar alcohol comprises
sorbitol.
11. The vaccine of claim 1, which does not contain an aluminum
compound.
12. The vaccine of claim 1, wherein the VLP comprises the
structural components of hepatitis virus, the immunogenic component
comprises an immunogenic component of human papilloma virus, and
the stabilizing agent comprises a sugar alcohol and/or a degraded
gelatin, and the vaccine does not contain an aluminum compound.
13. The vaccine of claim 1, which is a liquid.
14. The vaccine of claim 1, which is a lyophilized powder.
15. The vaccine of claim 1, which further contains a
pharmaceutically acceptable carrier selected from the group
consisting of alcohol, propylene glycol, fatty alcohols,
triglycerides, fatty acid esters, mineral oils, liquid petrolatum,
isopropylpalmitate, polyethylene ethanol, polyoxyethylene
monolauriater, sodium lauryl sulfate, an anti-oxidant, a humectant,
a viscosity stabilizer or modifier, a colorant or a flavoring
agent.
16. A method of treating or preventing an infection comprising
administering a therapeutically effective amount of the vaccine of
claim 1 to a mammal in need thereof, comprising determining the
therapeutically effective amount of the vaccine to be administered,
and administering that amount of the vaccine to the mammal.
17. The method of claim 16, wherein the vaccine is a lyophilized
powder and reconstituted to an aqueous or non-aqueous liquid prior
to administration to the patient.
18. The method of claim 16, wherein the vaccine is administered as
a liquid.
19. The method of claim 16, wherein administering is
intra-muscular, intra-peritoneal, or intra-venous.
20. The method of claim 16, wherein the patient is an infant, a
toddler, an adolescent, an adult or a senior.
21. The method of claim 16, wherein administration does not
generate a general inflammatory response or local inflammation at
the site of administration.
22. A method for the manufacture of a vaccine comprised of VLPs, an
immunogenic agent, and a stabilizing agent, comprising: generating
VLP comprising structural components of human hepatitis virus;
mixing the components with an immunogenic agent and a stabilizing
agent; and forming VLPs that encapsulate the immunogenic agent and
the stabilizing agent.
23. The method of claim 22, wherein structural components, the
immunogenic agent and the stabilizing agent are mixed at
approximately equimolar amounts.
24. The method of claim 22, wherein the VLPs that encapsulate the
immunogenic agent and the stabilizing agent are in an aqueous
mixture.
25. The method of claim 24, further comprising lyophilizing the
aqueous mixture.
Description
REFERENCE TO RELATED APPLICATIONS
[0001] This application claims priority to U.S. Provisional
Application No. 62/214,526 entitled "VLP Stabilized Vaccine
Compositions" filed Sep. 4, 2015, the entirety of which is
specifically incorporated by reference.
BACKGROUND
[0002] 1. Field of the Invention
[0003] The invention is directed to compositions and methods for
stabilizing biological materials and, in particular, vaccines. In
particular, the invention is directed to vaccines that are
encapsulated within VLPs that contain an immunogenic or active
component and a stabilizer. Preferred stabilizers include sugar
alcohols sorbitol and degraded gelatins. The invention further
relates to stabilized vaccines for the treatment or prevention of
infections cause by bacterial, viral, parasitic or other infections
and the manufacture of such vaccines.
[0004] 2. Description of the Background
[0005] HPV, or human papillomavirus, is a human DNA virus of the
papillomavirus family of viruses. Similar to other
papillomaviruses, HPVs infects keratinocytes of the skin and mucous
membranes. Typically, HPV infections are subclinical and cause
little to no physical symptoms. Infections can become clinical and
lead to the development of benign papilloma, also called warts or
squamous cell papilloma. Clinical infections can also become
cancerous developing into cancer of, for example, the throat and
reproductive tissues such as the cervix.
[0006] A large majority of clinical HPV infections regress to
subclinical in one to two years. In patients where the subclinical
infection persists, which occurs in five to ten percent of infected
women, there is high risk of developing precancerous lesions of the
vulva and cervix that progress to invasive cancer. As progression
from subclinical to clinical infection takes years, there are many
opportunities for detection and treatment of pre-cancerous
lesions.
[0007] In developed countries, cervical screening using a
Papanicolaou (Pap) test or liquid-based cytology is used to detect
abnormal cells that may develop into cancer. If abnormal cells are
found, women are invited to have a colposcopy during which biopsies
can be taken and abnormal areas removed. The removal of abnormal
cells is highly effective in preventing the development of cervical
cancer.
[0008] Two HPV vaccines are available, CERVARIX.TM. and
GARDASIL.TM.. Each prevents infection caused by HPV types 16 or 18.
These two strains are believed to be responsible for over seventy
percent of cervical cancers. As with all vaccines, HPV vaccines
need to be stabilized during transportation and storage and
therefore contain stabilizers. Stabilizers are chemical compounds
added to a vaccine formulation to enhance vaccine stability during
low temperature storage or storage post-lyophilization.
[0009] One such chemical stabilizer is referred to as SPGA and
contains 218 mM sucrose, 3.76 mM KH.sub.2PO.sub.4, 7.1 mM
K.sub.2HPO.sub.4, 4.9 mM potassium glutamate, and 1% serum albumin.
Modifications of SPGA include monosodium glutamate as a substitute
for monopotassium glutamate, starch hydrosylate such as glucose or
dextran as a substitute wholly or partly for sucrose, and casein or
PVP substituted wholly or partly for albumin. Another well-known
chemical stabilizer comprises approximately 3.5% hydrolyzed
gelatin, 3.5% sorbitol, 1.0% Medium 199, along with minimal amounts
of sodium bicarbonate and phenol red. Other stabilizers comprise
minute amounts of DPG solution, which contains, among other
compounds, cysteine, glutathionine, ascorbic acid, vitamin A and
USP. Additional processes and stabilizing agents are disclosed in
U.S. Pat. Nos. 3,985,615; 3,915,794; 4,000,256; 4,147,772;
4,537,769; 4,555,401; 4,849,358; 4,985,244; 5,139,776; 7,998,488;
8,142,795; 8,551,523; 8,557,253; 8,795,683 and International
Application Publication No. WO1998028000, each of which is
incorporated herein by reference.
[0010] Nevertheless, all of these stabilizers have shortcomings,
either an inability to maintain stability in higher temperatures or
fail after a period of time less than optimal. Thus, a need
currently exists for an effective stabilization chemical
formulation and/or process that maintains immunogenicity of the
vaccine and imparts no side effects.
SUMMARY OF THE INVENTION
[0011] In general, the invention is directed to compositions and
methods for stabilizing vaccine formulation and especially human
papilloma virus (HPV) vaccines.
[0012] One embodiment of the invention is directed to vaccines
comprising VLPs (virus like particles) that encapsulate an
immunogenic component and a stabilizing agent. The immunogenic
component comprises the immune-stimulating portion or active
portion of the vaccine that generates an immune response. Preferred
immune stimulating portions include immunogenic portions of an
infectious virus, bacterium or parasite. Preferred VLPs comprise
structural components of a virus such as, for example, the
structural components of hepatitis virus or human papilloma virus
(HPV), and VLP do not contain genetic material that permits
replication of particles. Preferably the stabilizing agent
comprises one or more gelatins, one or more amino acids, or one or
more sugar alcohols, or any combination thereof. Preferred gelatins
are degraded chemically or mechanically to fragments with an
average molecular weight of ten kilodaltons or less, or eight
kilodaltons or less. Also preferably, vaccines of the invention do
not require the addition of aluminum compounds such as aluminum
phosphate. Vaccines of the invention may be maintained for long
periods of time as a liquid, or even longer periods of time as a
lyophilized powder. Preferred vaccines further contain
pharmaceutically acceptable carriers such as, for example, an
alcohol, propylene glycol, fatty alcohols, triglycerides, fatty
acid esters, mineral oils, liquid petrolatum, isopropylpalmitate,
polyethylene ethanol, polyoxyethylene monolauriater, sodium lauryl
sulfate, an anti-oxidant, a humectant, a viscosity stabilizer or
modifier, a colorant or a flavoring agent.
[0013] Another embodiment of the invention is directed to methods
for the administration of stabilized vaccines of the invention to a
patient. Preferred patients are mammals that have a functioning
immune system. Methods comprise determining the therapeutically
effective amount of the vaccine to be administered, and
administering that amount of the vaccine to a patient in need
thereof. Preferably the vaccine is a lyophilized powder and
reconstituted to an aqueous or non-aqueous liquid prior to
administration to the patient. Also preferably the vaccine is
administered as a liquid and administering is intra-muscular,
intra-peritoneal, or intra-venous. Preferred patients include, but
are not limited to an infant, a toddler, an adolescent, an adult or
a senior. Preferably administration of a vaccine of the invention
does not generate a general inflammation response in the patient or
local inflammation at the site of administration.
[0014] Another embodiment of the invention is directed to methods
for the manufacture of stabilized vaccines of the invention
comprised of VLPs, an immunogenic agent, and a stabilizing agent,
comprising: generating VLP comprising structural components of
human hepatitis virus; mixing the components with an immunogenic
agent and a stabilizing agent; and forming VLPs that encapsulate
the immunogenic agent and the stabilizing agent. Preferably the
structural components, the immunogenic agent and the stabilizing
agent are mixed at approximately equimolar amounts. Also preferably
the VLPs that encapsulate the immunogenic agent and the stabilizing
agent are in an aqueous mixture. Preferably manufacturing
comprising lyophilizing the aqueous mixture. Also preferably, no
aluminum-containing compounds are added to the vaccine
composition.
[0015] Other embodiments and advantages of the invention are set
forth in part in the description, which follows, and in part, may
be obvious from this description, or may be learned from the
practice of the invention.
DESCRIPTION OF THE INVENTION
[0016] There are many vaccines that are currently available that
provide protection against viral and bacterial infections. Each of
these vaccines contains various non-active components that increase
stability and maintain the effectiveness of the vaccine over
periods of time.
[0017] It has been surprisingly discovered that vaccine stability
can be increased by containing the vaccine in a virus-like particle
(VLP). Preferably VLPs contain the immunogenic component of the
vaccine plus one or more stabilizers and one or more
pharmaceutically acceptable carriers. Preferred stabilizers include
sorbitol, amino acids, gelatin degraded to an average molecular
weight of 10,000 MW or less, or combinations thereof. Increased
stability includes an increased thermostability and an increased
stability over time. Although the increased stability herein is
stated in reference to HPV vaccines, as is believed to be clear to
those skilled in the art, the increased stability is equally
applicable to most any vaccine composition including vaccines to
prevent or treat viral, bacterial, parasitic and/or other
infections.
[0018] Virus-like particles are comprised of viral structural
proteins and resemble viruses, but are non-infectious and unable to
replicate as they contain no viral genetic material. The VLPs
self-assemble from expression of the viral structural proteins,
such as, for example, envelope or capsid proteins in vitro such as
for example in a test tube, and/or in vivo such as for example in
cell culture. VLPs of the invention are preferably derived from
hepatitis virus (e.g., Hepatitis B virus) and composed of the small
HBV derived surface antigen (HBsAg), but may also be derived from
the structural components of parvoviruses (e.g. adeno-associated
virus), retroviruses (e.g. HIV), vesicular stomatitis virus (e.g.,
VSV), hepatitis virus (e.g. hepatitis A, B, C, D or E virus),
flaviviruses (e.g., West Nile virus, dengue virus, tick-borne
encephalitis virus, yellow fever virus, Zika virus and several
other viruses that cause encephalitis), and combinations thereof.
VLPs can be produced in a variety of cell culture systems including
mammalian cell lines, insect cell lines, yeast cell lines, and
plant cell lines. VLPs may be formed in a buffered solution
containing one or more of chelating and/or reducing agents.
[0019] Accordingly, one embodiment of the invention is directed to
a vaccine containing an immunogenic agent and a stabilizing agent
encapsulated into a VLP. The immunogenic agent of the VLP comprises
an antigen or other structure that stimulates an immune response in
an individual. Immunogenic agents include, but are not limited to
peptides, proteins, lipids, fatty acids, polysaccharides,
lipopolysaccharides. Typically, immunogenic agents are surface
antigens of an infectious particle. Antigens may be specific for a
particular infectious agent or combination of agents such as, for
example, viral infectious agents (e.g., enterovirus, hepatitis
virus, human immunodeficiency virus {HIV}, human papilloma virus
{HPV}, influenza virus, pertussis virus, rubella virus, tetanus,
varicella virus {VZV}, flavivirus, West Nile virus, dengue virus,
tick-borne encephalitis virus, yellow fever virus, Zika virus),
bacterial infectious agents (e.g., chlamydia, clostridium,
diphtheria, meningococcal, streptococcal, staphylococcal,
pneumococcal), parasitic infectious agents (e.g., giardia, malaria
{plasmodium}), and/or an agent that causes sepsis or septicaemia.
Preferably the immune response is sufficient to protect the
individual from subsequent infections for a period of time.
Preferably the vaccine is effective and protects a patient from
infection for six months or greater, one year or greater, two years
or greater, five years of greater, or ten years or greater The
immune response generated in response to the immunogenic agent of
the invention generates a humoral immune response, a cellular
immune response, or preferably both in the individual. Preferred
cellular response include a T cell response, and/or a phagocytic
response, and also preferably a memory cell response.
[0020] Preferred stabilizing agents to include in VLPs of the
invention include, but are not limited to one or more of
carbohydrates such as glucose and/or fructose, sugar alcohols such
as glycerol, erythritol, threitol, avabitol, xylitol, ribitol,
mannitol, galactitol, fucitol, iditol, inositol, volemitol,
isomalt, lacitol, sorbitol, maltotriitol, maltotetraitol,
polyglyitol or dextrin, dehydrated sugar alcohols such as
isosorbide, amino acids such as arginine, proline, and/or lysine,
substituted amino acids such as hydroxyl-alanine, animal-derived
gelatins such as human collagen, BSA, and/or fish gelatin, and/or
plant-derived carbohydrates such as pectin and cellulose.
Preferably gelatins of the invention are degraded to an average
molecular weight of 100 kilodaltons (kD) or less, 50 kD or less, 20
kD or less, 10 kD or less, 8 kD or less, or 5 kD or less. Also
preferably, vaccines of the invention do not require the addition
of aluminum, magnesium or manganese compounds such as aluminum
phosphate or other forms of aluminum, magnesium or manganese, or
preferably any metal compound. Stabilizing agents may be added at
from 1-10% of the dry weight of the VLP components, or from 1-10%
of the dry weight of the VLP components plus the immunogenic agent,
or preferably from 2-4% of the dry weight of either composition.
Also preferably, the stabilizing agent is at a concentration of
from 1 mM to 50 mM of the aqueous mixture, preferably at a
concentration of from 10-20 mM.
[0021] Preferably the vaccine is maintained as a liquid, but the
liquid may be lyophilized and stored as a dry powder until use
wherein the powder is rehydrated in an appropriate carrying agent
(aqueous or non-aqueous) for administration to the individual.
Administration is preferable by injection which may be intra
peritoneal (i.p.), intra muscular (i.m.), and/or intra venous
(i.v.), or localized to the site of an infection.
[0022] Preferably the vaccines of the invention as liquids or
powders are stable at 4.degree. C. or greater, 15.degree. C. or
greater, 25.degree. C. or greater, 37.degree. C. or greater,
40.degree. C. or greater, 50.degree. C. or greater, or 100.degree.
C. or greater. Also preferably, the vaccines of the invention as
liquids or powers are stable at 15.degree. C. or less, more
preferably to 0.degree. C. or less, more preferably to minus
20.degree. C. or less, and more preferably to minus 50.degree. C.
or less. Also preferably, stability of the vaccines is maintained
for six month or greater, for eight months or greater or for twelve
months or greater. Preferably the vaccines of the invention are
stable through varying temperatures over time which may include
multiple freezing and thawing.
[0023] Another embodiment of the invention is directed to methods
for the administration of vaccines of the invention to patients in
need thereof for treating or preventing an infection. The method
comprises administering a therapeutically effective amount of the
vaccine of the invention to a mammal, comprising determining the
therapeutically effective amount of the vaccine to be administered.
The therapeutically effective amount is typically determined by
based on the weight of the mammal and the strength or
responsiveness of the patient's immune system and can be determined
by those skilled in the art. The therapeutically effective amount
is administered to a patient in need thereof, which may be to treat
an active or suspected infection or prevent an infection. The
vaccine may have been obtained from a lyophilized powder and
reconstituted to an aqueous or non-aqueous liquid prior to
administration to the patient. Preferably the vaccine is
administered as a liquid, which may be intra-muscular,
intra-peritoneal, or intra-venous, and the patient may be an
infant, a toddler, an adolescent, an adult or a senior.
Surprisingly, the vaccine of the invention does not generate side
effects such as redness or inflammation at the injection site, and
does not generate a generalized fever or inflammation, or other
unwanted side effects for the patient. Preferably an
immunologically effective vaccine contains only the fully formed
VLPs containing immunogenic and stabilizing agents, and nothing
further such as, for example, no added ionic or non-ionic
surfactants.
[0024] Another embodiment of the invention is directed to method
for the manufacture of vaccines of the invention. Structural
components of viruses are obtained by methods well known to those
skilled in the art and exclusive of any nucleic acid material that
would allow the components and resulting particles to replicate.
Predetermined molar amounts of the structural components are mixed,
preferably at room temperature or below, with approximately
equivalent molar amounts of one or more immunogenic agents and one
or more stabilizing agents of the invention, such that the VLPs
encapsulate the one or more immunogenic agents and the one or more
stabilizing agents in roughly equivalent amounts. The fully formed
VLPs are separated from unformed VLPs and free structural and other
materials preferably by filtration, centrifugation or another
method known to those skilled in the art, thereby creating fully
formed VLPs containing one more immunogens and one or more
stabilizing agents. The fully formed VLPs may be stored as an
aqueous (e.g., water or saline) or non-aqueous (e.g., oils, fatty
acids) mixture, or lyophilized and stored as a powder. Preferably
storage until use is without significant loss of immunogenic
activity and, for example, may be for one month or longer, four
months or longer, six months or longer, or more preferably one year
or longer and at ambient temperatures, whether as a liquid or a
powder. Storage of vaccine without loss of immunogenic activity may
also be at less than ambient temperature such as, for example, at
20.degree. C. or less, at 10.degree. C. or less, at 4.degree. C. or
less, or at 0.degree. C. or less.
[0025] Other embodiments and uses of the invention will be apparent
to those skilled in the art from consideration of the specification
and practice of the invention disclosed herein. All references
cited herein, including all publications, U.S. and foreign patents
and patent applications, are specifically and entirely incorporated
by reference. The term comprising, where ever used, is intended to
include the terms consisting and consisting essentially of.
Furthermore, the terms comprising, including, and containing are
not intended to be limiting. It is intended that the specification
and examples be considered exemplary only with the true scope and
spirit of the invention indicated by the following claims.
* * * * *